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Volume 11
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Analysis of the Short- and Long-Term Immune Response in BALB/c Mice Immunized with Total Naegleria fowleri Extract Co-Administered with Cholera Toxin

by
Mara Gutiérrez-Sánchez
1,
Maria de la Luz Ortega-Juárez
1,
María Maricela Carrasco-Yépez
2,
Rubén Armando Herrera-Ceja
1,
Itzel Berenice Rodríguez-Mera
2 and
Saúl Rojas-Hernández
1,*
1
Laboratorio de Inmunobiología Molecular y Celular, Sección de Estudios de Posgrado e Investigación, Escuela Superior de Medicina, Instituto Politécnico Nacional, Mexico City C. P. 11340, Mexico
2
Laboratorio de Microbiología, Grupo CyMA, Unidad de Investigación Interdisciplinaria en Ciencias de la Salud y la Educación, UNAM FES Iztacala, Universidad Nacional Autónoma de México, Tlalnepantla de Baz C. P. 54090, Mexico
*
Author to whom correspondence should be addressed.
Trop. Med. Infect. Dis. 2026, 11(1), 22; https://doi.org/10.3390/tropicalmed11010022
Submission received: 23 October 2025 / Revised: 7 January 2026 / Accepted: 9 January 2026 / Published: 12 January 2026
(This article belongs to the Special Issue Naegleria fowleri and Emerging Amoebic Infections)
Versions Notes

Abstract

Background: Naegleria fowleri is a free-living amoeba that inhabits warm freshwater and causes primary amoebic meningoencephalitis (PAM), a rapidly fatal infection with >95% mortality. Due to the lack of early diagnosis and effective therapy, preventive vaccination represents a promising strategy. Methods: This study evaluated short- and long-term immune protection in BALB/c mice (20 mice per group) immunized intranasally with total N. fowleri extract co-administered with cholera toxin (CT). Mice were challenged with a lethal dose of trophozoites either 24 h (short-term) or three months (long-term) after the fourth immunization; the latter group received a booster 24 h before challenge. Serum and nasal washes were analyzed for IgA and IgG antibodies by immunoblot, and lymphocyte subsets from nasal-associated lymphoid tissue (NALT) and nasal passages (NPs) were characterized by flow cytometry. Results: Immunization conferred complete (100%) survival in the 24 h group and 60% protection in the 3-month group, whereas all control mice died. Immunoblotting showed that IgA and IgG antibodies recognized major N. fowleri antigens of 37, 45, 48 and 19, 37, and 100 kDa, respectively. Flow cytometry revealed increased activated and memory B lymphocytes, dendritic cells, and expression of CCR10, integrin α4β1, and FcγRIIB receptors, particularly in the 24 h group. Conclusions: Intranasal immunization with N. fowleri extract plus CT elicited both systemic and mucosal immune responses capable of short- and long-term protection. These findings highlight the potential of this immunization strategy as a foundation for developing effective vaccines against PAM.
Keywords: Naegleria fowleri; immunization; cholera toxin; nasal passages; nasal-associated lymphoid tissue Naegleria fowleri; immunization; cholera toxin; nasal passages; nasal-associated lymphoid tissue

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MDPI and ACS Style

Gutiérrez-Sánchez, M.; Ortega-Juárez, M.d.l.L.; Carrasco-Yépez, M.M.; Herrera-Ceja, R.A.; Rodríguez-Mera, I.B.; Rojas-Hernández, S. Analysis of the Short- and Long-Term Immune Response in BALB/c Mice Immunized with Total Naegleria fowleri Extract Co-Administered with Cholera Toxin. Trop. Med. Infect. Dis. 2026, 11, 22. https://doi.org/10.3390/tropicalmed11010022

AMA Style

Gutiérrez-Sánchez M, Ortega-Juárez MdlL, Carrasco-Yépez MM, Herrera-Ceja RA, Rodríguez-Mera IB, Rojas-Hernández S. Analysis of the Short- and Long-Term Immune Response in BALB/c Mice Immunized with Total Naegleria fowleri Extract Co-Administered with Cholera Toxin. Tropical Medicine and Infectious Disease. 2026; 11(1):22. https://doi.org/10.3390/tropicalmed11010022

Chicago/Turabian Style

Gutiérrez-Sánchez, Mara, Maria de la Luz Ortega-Juárez, María Maricela Carrasco-Yépez, Rubén Armando Herrera-Ceja, Itzel Berenice Rodríguez-Mera, and Saúl Rojas-Hernández. 2026. "Analysis of the Short- and Long-Term Immune Response in BALB/c Mice Immunized with Total Naegleria fowleri Extract Co-Administered with Cholera Toxin" Tropical Medicine and Infectious Disease 11, no. 1: 22. https://doi.org/10.3390/tropicalmed11010022

APA Style

Gutiérrez-Sánchez, M., Ortega-Juárez, M. d. l. L., Carrasco-Yépez, M. M., Herrera-Ceja, R. A., Rodríguez-Mera, I. B., & Rojas-Hernández, S. (2026). Analysis of the Short- and Long-Term Immune Response in BALB/c Mice Immunized with Total Naegleria fowleri Extract Co-Administered with Cholera Toxin. Tropical Medicine and Infectious Disease, 11(1), 22. https://doi.org/10.3390/tropicalmed11010022

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